JP3006919B2 - 9-purinylphosphonic acid derivative - Google Patents

Abstract

This invention relates to novel purine nucleoside phosphorylase inhibitors, to the methods and intermediates for their preparation and to their use as immunosuppressants, antilymphoma, antileukemic, antiviral and antiprotozoal agents.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a novel purine nucleoside phosphorylase inhibitor, a process for producing the same, an intermediate for the production thereof, an immunosuppressant, an antilymphoma agent, an antileukemia agent, an antiviral agent and an antigenic animal. It relates to its use as an agent.

[0002]

2. Description of the Related Art Purine nucleoside phosphorylase (P
NP) converts guanine and hypoxanthine ribo- and deoxyribonucleosides under normal in vivo conditions
It catalyzes phosphorolytic cleavage to the corresponding sugar phosphate and guanine or hypoxanthine. P
In the absence of NP, the concentration of uric acid is very low, while certain nucleoside substrates of PNP, such as (dGuo)
Are increasing in plasma and urine. dGuo is toxic to lymphoblasts, but T cells are more affected than B cells. In fact, in patients with a genetically acquired PNP deletion, B cell immunoglobulin production is normal or even elevated, but these patients have leukopenia and no T lymphocyte function. It is either missing or very depressed. While uncontrolled deletion of PNPs is clearly undesirable, there are many cases where controlled suppression of the immune system, and especially of T cells, is highly desirable. These include, for example, treatment of T-cell leukocytes, suppression of host-graft response in organ transplant recipients, and treatment of gout. Applicants have discovered a family of 9-purinylphosphonic acid derivatives, which are potent inhibitors of PNP and are therefore useful as immunosuppressants.

[0003]

More particularly, the present invention relates to novel purinylphosphonic acid derivatives of the formula: ## STR00001 ## tautomeric forms thereof, and pharmaceutically acceptable salts thereof.

Embedded image

In the formula, R is

Embedded image Wherein Ar is a group wherein the CH ₂ moiety next to Ar is attached to one ring carbon atom of the R ₉ substituted phenyl, R ₉ substituted thiophene or R ₉ substituted furan moiety, and Z moiety adjacent to Ar Is the bridging moiety attached to the second ring carbon atom of the R ₉ substituted phenyl, R ₉ substituted thiophene or R ₉ substituted furan moiety. Z is a part of (a), (b), (c), (d), or (e), wherein (a) is

Embedded image And (b) is

Embedded image And (c) is

Embedded image (d) is -C≡C-, and (e) is

Embedded image Provided that when Z is a moiety (b), Ar is other than a furan or thiophene moiety, n is an integer of 1 to 5 or 0, and R ₁ is —OH Or —SH, R ₂ is H or —NH ₂ , and R ₃ is H,
-NH _2, -OH or -NH-NH _2, R ₄ is H, R 'or ₄ is H, OH, or F, or R ₄ and R'
₄ forms a keto moiety with the carbon atom bonded together, R ₅ is C ₁ - is ₆ or R _'5 is alkyl, R ₆ is C ₁
- ₆ alkyl or R 'is a _6, wherein R' is ₅ and R _'6 are H, each of R ₇ and R ₈ are H, F, or C ₁ - is ₄ alkyl, R ₉ is H _{, Cl, Br, C 1 -} 6 alkyl, C ₁ - ₆ alkoxy, OH, is a NH ₂ or CH _3, when Ar is furan or thiophene, that R ₉ is other than OH or NH ₂ As a condition, X and Y are H, F, or Cl, provided that when n is 0, both X and Y are H.

[0005] As used herein C ₁ - ₄ or C ₁ - The term ₆ alkyl, a straight chain, and branched-chain saturated lower aliphatic hydrocarbyl moieties, each having a 4 or up to 6 carbon atoms And includes methyl, ethyl, isopropyl, sec-butyl, n-butyl, t-butyl, pentyl and the like. C ₁ - ₆ alkoxy moiety thereof Est -
It is a tell derivative. The Ar moiety bridging its adjacent CH ₂ and Z moieties is an R ₉ -substituted phenyl, R ₉ -substituted furan or R ₉ -substituted thiophene moiety, where phenyl is the 1,2-, 1, It can be bridged in the 3- or 1,4-position, and each of furan and thiophene is through a ring carbon atom in the 2,3-, 2,4-, 2,5- or 3,4-position. Can be bridged. The R ₉ substituent can be mono or di-substituted, and the substitution can be made at any other available ring carbon atom. The tautomeric enol-keto form is the 6-form of the purine nucleus.
-Exist in position.

The expression pharmaceutically acceptable acid addition salt is intended to apply to any non-toxic organic or inorganic acid addition salt of a base compound of the formula 1. Examples of suitable salts forming inorganic acids include hydrochloric, hydrobromic, sulfuric and phosphoric acids, and acid metal salts such as sodium monohydrogen orthophosphate and potassium hydrogen sulfate. Examples of suitable salts forming organic acids include mono, di, and tricarboxylic acids. Examples of such acids are, for example, acetic acid, glycolic acid, lactic acid,
Pyruvic acid, malonic acid, succinic acid, glutaric acid, fumaric acid
Luic acid, malic acid, tartaric acid, citric acid, ascorbic acid, maleic acid, hydroxymaleic acid, benzoic acid,
They are hydroxybenzoic acid, phenylacetic acid, cinnamic acid, salicylic acid, and 2-phenoxybenzoic acid. Other organic acids which form suitable salts are sulfonic acids such as methanesulfonic acid and 2-
Hydroxyethanesulfonic acid. Either the mono or di acid salts can be formed, and such salts can exist in either hydrated or substantially anhydrous form. Acid salts are standard technology,
For example, the free base is dissolved in an aqueous solution or aqueous alcohol solution, or in a solvent containing another suitable acid and isolated by evaporating the solution, or the free base is dissolved in an organic solvent. By reacting,
In this case, the salt separation can be carried out directly or by concentration of the solution. In general, the acid addition salts of the compounds of this invention are crystalline materials, soluble in water and various hydrophilic organic solvents, have higher melting points and increased stability as compared to their free base forms. Is shown.

The preparation of the compounds of formula 1 generally involves the activation of (-CH ₂ -Ar-Z-)-substituted phosphones with 6-chloropurine (2) activated.
(3) and carried out by a condensation reaction in which the resulting Ia intermediate is converted to the appropriate R, R ₁ , R ₂ , R ₃ -substituted purine derivative of formula I. The general condensation reaction is shown in the following reaction scheme.

Embedded image

Wherein R ₁ , R ₂ , R ₃ , Ar, Z, R ₅ and R ₆
Is as defined above, with the proviso that when Z is part (a), R ′ _{4 which} is derived can be a silyl ether;
Is bromo, iodo or hydroxy.

If it is desired to carry out the condensation reaction of the 6-chloropurine intermediate (2) with phosphonoaryl (3), where Q is a halide, the condensation is slightly excess (about 10%).
6-Cl purine reactant (2) with a base such as sodium hydride (NaH), potassium carbonate (K ₂ CO ₃ ) or cesium fluoride (CsF)
(In an amount of about 2 equivalents) in a non-reactive solvent such as dimethylformamide (DMF), preferably by reacting in a temperature range of about 0 ° C to 60 ° C, preferably at room temperature for about 4 to 18 hours. Is done.

When Q is OH, R 'is preferably phenyl, but in the presence of P (R') ₃ , which also includes methyl and isopropyl, diethyl azodicarboxylate (DEAD) The condensation is carried out under more neutral conditions according to the Mitsunobu reaction, using 0 to 60 ° C. in a suitable non-reactive solvent.

Of course, in each case, when the Ar moiety has an R ₉ substituent which can be affected by the reaction conditions of these condensations (or modification of purine bases), such substituents are optional. It can be modified to eliminate unwanted side reactions, and is converted back to the desired form at the appropriate stage. For example, if R ₉ is OH,
An intermediate ester or ether derivative can be formed, and at an appropriate stage, the ester or ether can
Is hydrolyzed back to These principles are well understood by those skilled in the art and need not be described at length here.

In the special case where R ₄ , R ′ ₄ , X and Y in formula (3) are all hydrogen,

Embedded image Purine bromomethyl derivative of phosphonyl aryl
Condensation with (2) and hydrogenating the resulting product, preferably using hydrogen gas, in the presence of palladium on carbon (H ₂ -Pd / C) according to standard techniques.

R ₄ is H, R ' ₄ is OH or R ₄ and R' ₄ form a keto moiety (as defined above). In the case of the phosphonyl halide

Embedded image [In the formula

Embedded image Represents t-butyldimethylsilyl ether.] It is provided to use a silyl ether (preferably t-butyldimethylsilyl ether) to protect the hydroxy moiety of The silyl protecting group is removed by acid hydrolysis as described. If the resulting alcohol is to be oxidized, it can be oxidized to the desired ketone by use of a Swelln oxidation reaction. In practice, it is preferred to form the silyl ether prior to activating the reactant of Formula 3 as described below.

The Q-activated reactant of Formula 3 can be prepared by methods well known in the art, preferably reacting before the OH group (if any) is activated with either a halo or hydroxy moiety. Use the protected intermediate.

Preferably, the bromination is carried out with N-bromo-succinimide (NBS) or other suitable N-bromoamides in the presence of a catalytic amount of benzoyl peroxide, and the reaction is preferably carried out using CCl as solvent. ₄ Performed in (carbon tetrachloride). The preparation of the reactant of Formula 3 where Q is OH is based on CeAm
Conversion of benzyl bromide (7) to its acetate by reaction with NO ₃ (ceric ammonium nitrate) or subsequent hydrolysis of the acetate with a catalytic amount of sodium methoxide in methanol. Decomposition and can be performed directly from (6) by performing the reaction using standard procedures well known in the art.

Embedded image

Wherein Ar, Z, R ₅ and R ₆ are as previously defined, with the proviso that R ′ ₄ is silyl ether (instead of OH) and R ₉ is protected rather than OH. A hydroxyl group (if applicable).

Once the enrichment of the 6-chloropurine base (2) has been carried out and the compound of formula Ia has been prepared, the modification at the 8,6- and / or 2-position is carried out in a stepwise manner to give the compound of the formula The desired R ₁ , R ₂ , and R ₃ portions of I are made.

R ₅ and R ₆ are both H and R ₁ is OH
To prepare a compound of formula I is is of the formula Ia corresponding Hosufe - Tojiesuteru (i.e. R ₅ and R ₆ are alkyl) are successively trimethylsilyl bromide in CH ₂ Cl ₂ (TMSBr), in acetonitrile Water and finally HCl (I
The reaction is carried out continuously at 90 ° C. in N). The compound of Ia is directly subjected to HCl / H ₂ O hydrolysis at 90 ° C. to make monoesters (R ₅ is H, R ₆ is alkyl, and R ₁ is OH).

To prepare compounds of formula I wherein R ₁ is SH, a compound of formula Ia is reacted with thiourea in acetic acid. Deesterification of the resulting 6-SH product by treatment with TMSBr and hydrolysis results in compounds where R ₁ is SH and R ₅ and R ₆ are H.

R ₁ is SH, R ₂ and R ₃ are as defined (Formula I) and R ′ ₄ is H or silyl ether (—O-SiMe
₃ ) In order to prepare a compound of formula I which is
The H analog is reacted with dimer phosphorus pentasulfide. R ' ₄
The resulting compound in which is a silyl ether can be converted to the alcohol, optionally oxidizing the alcohol to its keto analog by the methods described herein. Using the same reactants (where R ₃ is other than H), a brominating or iodinating agent such as bromine, N-bromo or N-iodimide (eg, 1,3-dibromo-5,5- Dimethyl-hydantoin, 1,
Halogenation (8- of the purine moiety) with 3-diodo-5,5-dimethylhydantoin, N-iodo-acetamide, preferably NBS or NIS, and most preferably N-bromoacetamide (NBA). Position), R ₃ is NH ₃ or —N
Can be produced a compound of Formula 1 is H-NH _2.
The 8-halo analog so prepared can be prepared in a suitable solvent such as water, ether, THF, p-dioxane, lower alkanol, ethylene glycol, chlorinated hydrocarbons (CCl ₄ , CH ₂ Cl ₂ ), DMF, HMPA or DMS
In O), it is reacted with hydrazine at a temperature of about 50 ° C. to 100 ° C., preferably with a 2-3 fold excess of hydrazine, although not necessary. Corresponding 8-NH ₂ compound, Raney hydrazine - built by reduction with nickel. The 8-halo analog used in the reaction with hydrazine is a reaction of an 8-halo compound with an alkali metal or alkaline earth metal salt of a benzyl alcohol (eg, benzyl alcohol), followed by intermediate formation Noble metal catalysts (eg Pt
/ C) can be used to produce compounds wherein R ₃ is OH by reduction with hydrogen gas at atmospheric pressure in the presence of / C).

In order to more easily show another route to make compounds of subtype (a) in which R ₄ is H and R ′ ₄ is OH or R ₄ and R ′ ₄ form a defined keto moiety. In order to clarify the concept regarding the synthetic route, the following reaction route is drawn.

Embedded image In the formula

Embedded image Represents t-butyldimethylsilyl, and Ar, X, Y, Bu
₄ NF, TMSBr are as defined above.

The preparation of the allyl phosphonate of formula 6 can be carried out using standard methods and techniques also known in the art. The particular route of synthesis will, of course, depend on the definition of the Z moiety.

An intermediate of formula 6 wherein Z represents subtype (a), ie

Embedded image Wherein Ar, R ₄ , R ′ ₄ , X, Y, R ₅ and R ₆ are as defined in formula I. If it is desired to produce the compound of formula (I), the particular synthetic route will be primarily R ₄ , R ′
₄ , depending on the specific definition of X and Y. When R ₄ and R ′ ₄ are hydrogen, the procedure is represented as follows:

Embedded image

Where M is Li, Na, -ZnBr, MgBr.
(Preferably lithium) and X and Y are H, F or C
l. In this reaction, the appropriate phosphonate is prepared by reaction with lithium diisopropylamide (LDA) or butyllithium in an inert solvent (eg, THF) at about -78 ° C. under an inert atmosphere (argon). The lithio derivative (17) to be reacted is reacted for 10 to 20 hours.
It is condensed with rubromide (16) and the reaction is stopped with a saturated aqueous ammonium chloride solution (NH ₄ Cl). M is -ZnBr
The condensation of the reactants (16 and 17) is preferably carried out at 20 ° C. in the presence of a catalytic amount of copper bromide. Zn and Mg bromides derivatives can also be made by standard methods.

If R ₄ is H and R ′ ₄ is OH or F, or R ₄ and R ′ ₄ form a keto moiety (as defined), the compound is as illustrated next. Built in.

Embedded image

The reaction of the aldehyde (18) with the lithio derivative (19) is carried out in THF at -78 ° C under argon for about 3 hours, and the reaction is then carried out at -78 ° C to -30 ° C with saturated aqueous NH ₄ Cl. And produce compound (20). Compound (20) as D
When treated with AST (diethylaminosulfur tetrafluoride), the reaction is carried out in dichloromethane at about 0 ° C. for about 15 to 25 hours and the reaction is stopped with an excess of methanol. The alcohols (20) are also converted to their keto forms using oxalyl chloride in DMSO, by using swelling oxidation reactions, or by using tetrapropylammonium partruthenate and N-methylmorpholine N-oxide. Oxidized. Alternatively, the ketone is of the formula wherein X "is chloro or alkoxy

Embedded image (19) wherein M is Li or -ZnB
It can be formed directly from the reaction of r).

In carrying out this latter reaction, which directly produces ketones, when X and Y are both H,
When X ″ is alkoxy (eg, methoxy), M is lithium, and X and Y are F, X ″ is chloro and M
Is particularly preferably -ZnBr.

When R ₄ is H and R ′ ₄ is OH, or R ₄ and R ′ ₄ form a defined ketone,
As described above for the condensation of reactants (2) and (3), the silyl ether of compound (20) is preferably used (eg a t-butyldimethylsilyloxy derivative). These silyl derivatives are made by reacting compound (19) with t-butyldimethylsilyl chloride in the presence of imidazole and activating the resulting product with NBS. Stage selective removal of at silyl protecting group of the latter, as shown by specified above, is carried out by treatment with tetrabutylammonium fluoride (Bu ₄ NF).

R ₄ and R ′ ₄ form a defined ketone;
In the particular case of preparing compounds of formula Ia where X and Y are H, it is preferred to use the following reaction scheme.

Embedded image

By condensation wherein Q in formula 3 is OH and R ₄ , R ' ₄ , X and Y are each H, Z is a subtype (a) of formula Ia
If it is desired to produce a compound of the general formula 3
It should be noted that it is preferable to reduce the intermediate of subtype (c) in order to obtain a suitable reactant falling within the range.

An intermediate wherein Z is included within the scope of formula 6 of subtype (b), ie

Embedded image If it is desired to prepare a compound of the formula wherein Ar 'is other than a furan or thiophene bridging moiety, standard procedures known in the art as analogous may be used. Generally, intermediates are prepared according to the following reaction scheme.

Embedded image Wherein R ' ₉ is as defined for R ₉ in formula I except OH, Q' is iodo, bromo, tosylate, mesylate, or trifluorolate leaving group; Is an integer of 1 to 5. The condensation is carried out with a base (eg, NaH, K ₂ CO ₃ or K
H) in the presence of a non-aqueous solvent such as DMF,
(THF or DMSO) using standard procedures well known in the art. In the particular case where n is 2, the cresol (ie, o-, n-, or p-cresol) is K
Reaction with ethylene carbonate in the presence of F gives benzoylethyl-1-ol-ether, which is converted to bromine in the presence of a base in benzene in the presence of triphenylphosphine (Pφ ₃ ). To give the benzoylethyl bromide, which is reacted with a lithio derivative of formula (17) by standard procedures to produce compound (30a) where n is 2.

Of course, when n is 0 and X and Y are both H, it is preferred to use the method of Q 'tosylate (compound 29) using NaH as the base in DMF.

A compound of formula 6 wherein Z represents subtype (c),

Embedded image If it is desired to prepare a compound of the formula wherein Ar, X, Y, R ₅ and R ₆ are as defined in formula I, the compound may be prepared by analogous methods known in the art. Can be manufactured.

When Y is H and X is H, F or Cl, aryl aldehyde (32) is

Embedded image Is condensed with the lithio derivative of the X-substituted diphosphonate derivative (33).

The reaction is carried out at -78 ° C. in THF, warming the mixture to about 20 ° C. before stopping and then adding saturated aqueous NH ₄
Hydrolysis with Cl. When Y is F and X is H, F or Cl, the preparation is carried out using a compound of subtype (a) (ie compound (35) wherein X 'is F or Cl), which is a base, preferably Is treated with DBU of tBuOK, DMAP in a non-reactive solvent (DMF or DMSO) at a temperature of about 40 ° C to 80 ° C. In this reaction a double bond is obtained by disappearance of HX '. By selecting the appropriate analog of compound (35) and following either treatment of the pregroup with a base, the desired Y and X compounds (34a) can be prepared when either HF or HCl is evolved. . The reaction is depicted as follows.

Embedded image

An intermediate of formula 6 wherein Z represents subtype (d), ie a formula

Embedded image If it is desired to prepare the compound of formula (I), the compound of formula (24) is treated with 2 equivalents of DAST at 0 ° C. to 20 ° C. in CH ₂ Cl ₂ for 1 to 5 hours and the reaction is To give the desired product 36.

Z is a compound of formula 6 of subtype (e), ie

Embedded image If it is desired to prepare a compound of the formula

Embedded image And can be carried out in a one- or two-stage olefination process.

In the one-step method, the formula

Embedded image Wittig type olefination process using a phosphonium ylide of the compound of formula 38 (especially X and Y are F)
Is reacted. The reaction is carried out at -78 ° C to 0 ° C in THF to generate a double bond. Alternatively, reactant 39 is
formula

Embedded image [Where Het is -Sφ, SiMe ₃ , Seφ, -SMe, or S
eMe] when reacted to give a compound of formula 41 when reacted.

Embedded image

[0039] When Het is SiMe _3, the peak - data - and Son olefination is used, the (a) 41 in DMF 0 ° C. ~
Treating with NAH at 60 ° C. or (b) 41 with an acid such as
Using high temperature treatment with PTSA (p-toluenesulfonsan). When Het is other than SiMe ₃ is, SOCl _2, POC 41 at a temperature of about -20 ° C. to 20 ° C. in CH ₂ Cl ₂
treated with l ₃ or PI ₃ and Et ₃ N or pyridine.

The following examples illustrate how the compounds of the present invention can be prepared.

EXAMPLES Synthesis of 1 : [2- [2-[(2-amino-1,6-dihydro-
Preparation of 6-oxo-9H-purin-9-yl) -methyl] phenyl] -1,1-difluoroethyl] phosphonic acid 1A : [2- [2-methylphenyl] -1,1-difluoroethyl] phosphonic acid Diethyl ester 30m dissolved in 30ml anhydrous tetrahydrofuran (THF)
mol (5.64 g) of diethyldifluoromethanephosphonate was slowly added to (LDA) lithium diisopropylamide (30
30 mmol and 31 mmol of diisopropylamine in ml of anhydrous THF
(Prepared at 0 ° C. from n-butyllithium) at −78 ° C. under argon. 45 mmol after 30 minutes
(8.33 g) of 2-bromo-o-xylene was added to the reaction mixture, the reaction mixture was stirred at -78 ° C for 15 hours and quenched by adding 20 ml of an aqueous saturated solution of ammonium chloride.
The crude mixture was evaporated to dryness and the residue was suspended in 50 ml of water and extracted three times with 100 ethyl acetate. The organic layer was dried over sodium sulfate, evaporated and filtered to give approximately 8 g of crude product, which was purified by flash chromatography on silica gel to give 3.6 g of 1A (41% yield) .

Preparation of 1B : [2- [2-[(2-amino-1,6-dihydro-6-chloro-9H-purin-9-yl) -methyl] phenyl]-
1,1-Difluoroethyl] phosphonic acid diethyl ester N-bromosuccinimide (3 mmol, 0.53 g) and benzoyl peroxide (5 mg) were added to a solution of 1A (20 ml in carbon tetrachloride).
3 mmol, 0.88 g). The mixture was refluxed by heating on a lamp for 90 minutes until all the succinimide appeared. The reaction mixture was filtered to remove succinimide and the filtrate was evaporated to dryness to give 1.1 g of an oil, which was then added to a stirred solution of the sodium salt of 6-chloroguanine (3.2 mmol).
Sodium hydride to 3.9 mmol of 6-chloroguanine
(Prepared in 5 ml DMF by addition at 0 ° C. under argon)). The reaction mixture was stirred at 20 ° C. for 20 hours, evaporated under reduced pressure and purified by flash chromatography on silica gel to give 1.15 g of the expected 1B (42%).

Preparation of 1 : [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl) -methyl] phenyl] -1,1
[Difluoroethyl] phosphonic acid 4 mmol (0.5 ml) of trimethylsilyl bromide (TMSBr) in 5
To a stirred solution of 1C (0.6 g, 1.3 mmol) in ml of anhydrous dichloromethane at 20 ° C. under argon. The reaction mixture was stirred for 20 minutes and 0.5 ml of TMSBr was added to the reaction mixture. After 20 hours, the reaction mixture was evaporated and the residue was dissolved in 3 ml of acetonitrile and quenched with approximately 0.2 ml of water.

The mixture was evaporated and the residue was treated with 7 ml of 1N HCl
Heated at 100 ° C. for 20 hours. The mixture was evaporated and the product was obtained after recrystallization twice from hot water: 200 mg (3
In 8% yield, the mother liquor contained essentially pure product for further isolation).

Synthesis of 2: [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl) -methyl] phenyl] -1-
Preparation of [fluoroethenyl] phosphonic acid 2A : [2- (2-methylphenyl] -1-fluoroethenyl] phosphonic acid diethyl ester 20 mmol (6.27 g) of bis (diethylphosphonyl) dissolved in 20 ml of anhydrous THF Slowly add fluoromethane slowly to lithium diisopropylamide (22 mmol n-butyllithium
0 ° C. to a solution of 22 mmol of diisopropylamine in THF
Prepared at -78 ° C. Three
After 0 min at −78 ° C. a solution of 30 mmol (3.5 ml) of freshly distilled o-tolualdehyde in 20 ml of THF is added to the reaction mixture, the reaction mixture is stirred at −78 ° C. for 3 h and at 20 ° C. Stirred for 5 hours, quenched with 20 ml of aqueous saturated ammonium chloride and evaporated to dryness. The residue was suspended in 30 ml of water and extracted three times with 100 ml of ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate, filtered, and evaporated to give 5 g of a crude material, which was purified by flash chromatography on silica gel to give 70 mmol (50%) of 2A. .

Preparation of 2D : [2- [2-[(2-amino-1,6-dihydro-6-chloro-9H-purin-9-yl) -methyl] phenyl] -1
[Fluoroethenyl] phosphonic acid diethyl ester Benzoyl peroxide (10 mg) in 15 ml of anhydrous carbon tetrachloride NB
Added to a suspension of S (10 mmol) and 2A (10 mmol). The mixture was heated to reflux with a lamp until all solids came up. The reaction mixture was filtered and evaporated to give 2C as an oil, which was then dissolved in 4 ml of anhydrous DMF and (10 mmol of NaH (60% solution W
/ V) to 10 mmol of 6-chloro-guanine in 10 ml of anhydrous DMF.
(Prepared by addition at 0 ° C. under argon) to a stirred solution of the sodium salt of 6-chloro-guanine. 20
After 20 h at C the reaction mixture was evaporated to dryness and the crude residue was purified directly by flash chromatography on silica gel to give 4 mmol of the product 2D (40% yield).

Preparation of 2 : [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl) -methyl] phenyl] -1-
The preparation of 2 from fluoroethenyl] phosphonic acid 2D was performed using the procedure previously described for converting 1C to 1.

Synthesis of 3 : [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl) -methyl] -phenyl] -1,
Preparation of 1,2-trifluoroethyl] phosphonic acid 3A: [2-hydroxy-2- (2-methylphenyl) -1-
Difluoroethyl] phosphonic acid diethyl ester 42.5 mmol of diethylphosphinyldifluoromethane (8 g) in 42 ml of anhydrous THF at −78 ° C. under argon at −78 ° C. in freshly prepared lithium diisopropylamide (42.5 m
mol) was slowly added. -78 reaction mixture
Stir at 35 ° C. for 35 minutes and dissolve in 7.65 g of 42 ml of THF
g of o-tolualdehyde (63.75 mmol) are added to the reaction mixture, the reaction mixture is stirred at -78 ° C for 4 hours, quenched at -78 ° C by adding 40 ml of saturated aqueous ammonium chloride and evaporated under reduced pressure I let it. The residue is suspended in water, 20
Extracted three times with 0 ml of ethyl acetate. The organic layer was washed with brine, dried over sodium sulfate, filtered, evaporated and purified by flash chromatography on silica gel.
0.67 g of 3A was provided as white crystals (81% yield).

Preparation of 3B : [2-Fluoro-2- [2-methylphenyl] -1,1-difluoroethyl] phosphonic acid diethyl ester To a stirred solution of 3A (4.6 g, 15 mmol) in 20 ml of anhydrous dichloromethane was added 2.3. 20 ml of diethylaminosulfur trifluoride (DAST)
C. was added dropwise at 0.degree. C. under argon. After 2 hours at 20 ° C., the reaction mixture was slowly quenched at 0 ° C. with excess methanol (5 ml),
Evaporated to dryness and purified directly by flash chromatography on silica gel to give 4.22 g of product 3B (91%) yield.

Preparation of 3C: [2- [2-[(2-amino-1,6-dihydro-6-chloro-9H-purin-9-yl) -methyl] phenyl]-
The bromination reaction of 1,1,2-trifluoroethyl] phosphonic acid diethyl ester 3B followed by condensation with 6-chloroguanine was performed as described for preparing 1C from 1B.

Preparation of 3: [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl) -methyl] phenyl] -1,
1,2-Trifluoroethyl] phosphonic acid Final product 3 was isolated after deprotection with TMSBr / CH ₂ Cl ₂ and 1N HCl in water as described for preparing 1 from 1C. .

Synthesis of 4: [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl) -methyl] -phenyl] -2-
Preparation of hydroxy-1,1-difluoroethyl] phosphonic acid 4A: [2- (t-butyldimethylsilyloxy) -2 (2-
Methyl-phenyl) -1,1-difluoroethyl] phosphonic acid diethyl ester Diethylphosphinyldifluoromethane (4.7 g, 25 mmol) dissolved in 25 ml of anhydrous THF was charged under argon with lithium isopropylamide (LDA) (25 mmol of n -Butyl lithium 2
(Prepared by reacting 5 mmol of diisopropylamine with 25 ml of THF at 0 ° C.) at −78 ° C. -
After 35 minutes at 78 ° C., o-tolualdehyde (3
(0 mmol, 3.6 g) was added to the reaction mixture. After 3 hours at −78 ° C., 30 mmol of t-butyldimethylsilyl chloride are added to the reaction mixture, and the reaction mixture is stirred at −20 ° C. for 2 hours,
Stopped with 1 liter of water, evaporated and extracted with three 120 ml portions of ethyl acetate. The organic layers were collected, dried over sodium sulfate, filtered, evaporated and purified by flash chromatography on silica gel to give 8.8 g of product 4A. (2
1 mmol, 84% yield).

Preparation of 4B: [2- (2-amino-1,6-dihydro-
6-oxo-9H-purin-9-yl) -methyl] phenyl]-[2- (t-
Butyldimethylsilyloxy) -1,1-difluoroethyl]
The preparation of 4B from phosphonic acid diethyl ester 4A was performed using the procedure described for converting 1A to 1C.

Preparation of 4C: [2-[(2-amino-1,6-dihydro
-6-oxo-9H-purin-9-yl) -methyl] phenyl] -2-hydroxy-1,1-difluoroethyl] phosphonic acid diethyl ester 12 mmol (3.85 g) of tetrabutylammonium fluoride in 150 ml at a time Added to a stirred solution of 4B (6 mmol) dissolved in THF. The reaction mixture was stirred at 20 ° C. for 20 hours, evaporated to dryness and purified by flash chromatography on silica gel to give 4.8 mmol of 4C (80% yield).

Preparation of 4: [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl) -methyl] phenyl] -2-
Compound 4 was obtained from hydroxy-1,1-difluoroethyl] phosphonic acid 4C by two previously described chemical deprotection steps (TMSBr: H ₃ O ⁺ ).

Synthesis of 5: [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl) -methyl] -phenyl] -2,
Preparation of 2-dihydroxy-1,1-difluoroethylphosphonic acid 5A from 4C: [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl)- Methyl] -phenyl] -2-
Hydroxy-1,1-difluoroethylphosphonic acid diethyl ester 60 mmol (4.3 mmol) dissolved in 25 ml of anhydrous dichloromethane
l) DMSO was added to a stirred solution of 30 mmol of oxalyl chloride (26 ml) in 25 ml of anhydrous dichloromethane under argon.
It was added dropwise at ° C. The reaction mixture was stirred at -65 ° C for 5 minutes, 25m
20 mmol of 4C dissolved in L CH ₂ Cl ₂ was added. The reaction flask was removed from the cooling bath for several minutes and the mixture was again
Stirred at 15 ° C. for 15 minutes. At that time 100 mmol (13.8 ml) of triethylamine was added to the reaction mixture and the reaction mixture was
Stir at −65 ° C. for 10 minutes, stop with aqueous citric acid, and for several minutes
Stir at 20 ° C., extract with dichloromethane (3 × 75 ml portions), wash with brine, dry over Na ₂ SO ₄ , filter, evaporate, purify by flash chromatography on silica gel, purify 13.5 m Molar product 4C was obtained. (67%).

Preparation of 5: [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl) -methyl] -phenyl] -2,
2-Dihydroxy-1,1-difluoroethyl] phosphonic acid The final product 5 was obtained from 5A by two deprotection steps performed to produce 1 from 1C.

Synthesis of 6: [3- [2-[(2-amino-1,6-dihydrido)
B-6-chloro-9H-purin-9-yl) -methyl] -phenoxy]-
1,1-difluoropropyl] phosphonic acidPreparation of 6A: 3- (2-methylphenoxy) -1-propanol  300 mmol o-cresol, 334 mmol ethylene carbonate
And 325 mmol of potassium fluoride in 100 ml of anhydrous D
The mixture was added to MF and stirred at 125 ° C. for 50 hours under argon. 40m
Of ethylene carbonate and 40 mmol of KF
And the reaction mixture was stirred at 125 ° C for a further 24 hours.
Was. The reaction mixture was cooled to 20 ° C., filtered and evaporated. Remaining
The distillate is flash chromatographed on silica gel.
Purification afforded 38.6 g of the expected product (85% yield).

[0058]Production of 6B: 3- (2-methylphenoxy) -1-
Bromopropane  10 dissolved in 30 ml of benzene (or acetonitrile)
g bromine (62.5 mmol) slowly into 100 ml benzene
(Or triphenylphosphine in acetonitrile)
(64 mmol) was added to the stirred solution. 15 minutes later, 35 ml of Ben
Triethyl alcohol dissolved in zen (or acetonitrile)
Min (64 mmol) was added to the reaction mixture, followed by a 50 ml bottle.
Starting material 6A dissolved in benzene (or acetonitrile)
(9.68 g, 63.7 mmol) was added. 20 reaction mixture
Stir at 20 ° C for 20 minutes.
Filter (to remove sid) and evaporate to give silica gel.
9.8 g purified by flash chromatography on a
The expected product is obtained.

[0059]Production of 6C: [3- (2-methylphenoxy) -1,
1-Difluoropropyl] phosphonic acid diethyl ester  30 mmol (5.64 g) dissolved in 30 ml of anhydrous THF
Difluoromethyl O, O-diethyl phosphonate slowly
And 37 mmol of LDA (30 ml of TH
31 mmol of n-butyllithium and 31 mmol of diisopropane in F
(Prepared from ropylamine). Reaction mixing
The mixture was stirred at -78 ° C for 30 minutes, dissolved in 10 ml of anhydrous THF.
The starting material (6B, 20 mmol) was added to the reaction mixture.
Stirring is continued at -78 ° C for 3 hours and the temperature is slowly raised to 20 ° C.
And stop the reaction mixture with saturated aqueous ammonium chloride
did. The crude mixture is then evaporated and extracted with ethyl acetate
Was. Collect the organic layer, wash with water and brine, and add sodium sulfate
Dried on a filter, filtered, evaporated and dried over silica gel.
Purified by flash chromatography, 40% (16 mmol)
The expected condensation product was obtained.

[0060]Production of 6D: [3- [2-[(2-amino-1,6-dihi
Dro-6-chloro-9H-purin-9-yl) -methyl] -phenoki
[S] -1,1-difluoropropyl] phosphonic acid diethyles
Tell  6 m of starting material 6C dissolved in 15 ml of anhydrous carbon tetrachloride
6 mmol of N-bromosuccinimide and several mg of peroxide
Heat with lamp with benzoyl for 35 minutes. Crude mixture
To remove succinimide, and evaporate the filtrate to dryness.
And dissolved in 8 ml of anhydrous DMF at 20 ° C. under argon
6.5 mmol 6-chloroguanine and 13 mmol potassium carbonate
For 24 hours. The mixture was evaporated to dryness and the residue
In 50 ml of ethyl acetate, ammonium chloride and salt
Wash with water, dry over sodium sulfate, filter and evaporate
And flash chromatography on silica gel.
Purification gave 3 mmol of the expected product.

[0061]Preparation of 6: [3- [2-[(2-amino-1,6-dihydrid
B-6-chloro-9H-purin-9-yl) -methyl] -phenoxy]-
1,1-difluoropropyl] phosphonic acid  9 mmol of freshly distilled TMSBr is slowly added to 10 ml
Starting material 6D dissolved in anhydrous dichloromethane (3m
To a stirred solution at 20 ° C. under argon. Reaction mixing
The material was stirred at 20 ° C. for 20 hours and evaporated to dryness. 8 ml of residue
Dissolve in anhydrous acetonitrile and stop with 10 mmol water
I let it. A white precipitate formed, which was separated by filtration and collected.
Gives the expected product and can be used without further purification
Used in stages.

2 mmol of the starting material dissolved in 10 ml of 1N HCl and 2 ml of THF were heated at 90-100 ° C. for 20 hours.
The reaction mixture was cooled to 20 ° C., evaporated to dryness, dissolved in saturated aqueous triethylammonium bicarbonate, filtered and 1N H 2
Crystallized by addition of Cl.

The white solid was collected, dried under reduced pressure and
The mole of expected product was obtained as the hemihydrate.

[0064]Synthesis of 7: [2-[(2-amino-1,6-dihydro-6
-Oxo-9H-purin-9-yl) -methyl] -phenoxy] -methyl
Ruphosphonic acid Preparation of 7A: 2-Methylphenoxymethylphosphonic acid die
Chill ester  10 ml of sodium hydride (8 mmol of a 60% suspension in oil)
Stirring of o-cresol (8 mmol, 864 mg) in anhydrous DMF
The solution was added at 20 ° C. under argon. 45 minutes later, 3 ml DMF
O, O-diethylmethylphosphonate tosylate dissolved in
(8 mmol, 2.54 g) was added to the reaction mixture.
Is stirred at 60 ° C. for 20 hours and evaporated under reduced pressure, silica gel
Purified by flash chromatography above, 1.1 g of raw
The product was obtained (69% yield).

[0065]Preparation of 7B: [2-[(2-amino-1,6-dihydro
-6-Chloro-9H-purin-9-yl) -methyl] -phenoxy] -meth
Tylphosphonic acid diethyl ester  10ml CCl_Four7A phosphonate (1.03g, 4m
), N-bromosuccinimide (4.2 mmol, 743 mg) and
And several milligrams of benzoyl peroxide under a heating lamp
Heated to reflux. After 35 minutes, the reaction mixture was filtered and evaporated.
To give 1.3 g of oil, which is dissolved in 3 ml of anhydrous DMF
6-chloroguanine (4.4mM) in 6ml anhydrous DMF
Of potassium carbonate (10 mmol, 1.38 g)
The suspension was added at 20 ° C. under argon. After 40 hours, mix the reaction
The compound was evaporated under reduced pressure and flash chromatographed on silica gel.
Purify by chromatography and obtain 1.25 g of the expected product
(74% yield).

[0066]Preparation of 7: [2-[(2-amino-1,6-dihydro-6
-Oxo-9H-purin-9-yl) -methyl] -phenoxy] -methyl
Ruphosphonic acid  Chemical deprotection using TMSBr as previously described,
An aqueous hydrolysis is then performed to give the title compound.

[0067]Synthesis of 8: [2- [2-[(2-amino-1,6-dihydrid
B-6-oxo-9H-purin-9-yl) -methyl] -phenyl] -1,
1-Difluoro-2-propenyl] -phosphonic acid Preparation of 8A: [2- [2-methylphenyl] -1,1-difluoroo
Xoethyl] -phospho Acid diethyl ester  50m solution dissolved in 50ml dimethoxyethane (DME)
(13.35 g) of O, O-diethylbromodifluoromethane
The rate at which the sulphonate is maintained at a slow and gentle reflux
Freshly activated zinc in 55 ml of DME (55 mmol)
Was added to the stirred suspension. Stir the reaction mixture at 20 ° C for 2 hours
60 mmol of o-toluic acid dissolved in 15 ml of DME
Chloride (25 g) is added to the reaction mixture, which is
While stirring. Filter the crude material over celite and evaporate the filtrate.
Evaporate to dryness and flash chromatography on silica gel
And purified directly by 30-mole to give 30 mmol of product 8A (60
%).

[0068]Production of 8B: [2- [2-methylphenyl] -1,1-
Difluoro-2-propenyl] -phosphonic acid diethyl ester
Le  35 mmol of n-butyllithium (1.6 N solution in hexane 21.
8 ml) in 35 ml of methyltriphenyl in 50 ml of THF
The stirred suspension of phosphonium bromide was
Added below. The reaction mixture is stirred at 0 ° C. for 2 hours, and
30 mmol of compound 8A dissolved in 30 ml of THF was added to -78.
C. was added to the reaction mixture. 2 hours at -78 ° C and at 0 ° C
After stirring for 2 hours, the reaction mixture was washed with saturated aqueous ammonium chloride.
And hydrolyzed. The crude product was evaporated under reduced pressure to give 100 ml
Extracted three times with ethyl acetate. Normal work-up and
Purification by flash chromatography on silica gel
Yielded 18 mmol of 8B (60% yield).

[0069]Production of 8C: [2- [2-[(2-amino-1,6-dihi
Dro-6-chloro-9H-purin-9-yl) -methyl] -phenyl]-
1,1-difluoro-2-propenyl] -phosphonic acid diethyl ester
Steal  Bromination reaction of 8B and subsequent condensation with 6-chloroguanine
As described for producing 1C from 1B
Carried out.

[0070]Preparation of 8: [2- [2-[(2-amino-1,6-dihydrid
B-6-oxo-9H-purin-9-yl) -methyl] -phenyl] -1,
1-Difluoro-2-propenyl] -phosphonic acid  Final product 8 as described for preparing 1 from 1C
And TMSBr / CH_TwoCl_TwoAnd 1N HCl in water
After protection, it was isolated from 8C.

[0071]Synthesis of 9: [2- [2-[(2-amino-1,6-dihydrid
B-6-oxo-9H-purin-9-yl) -methyl] -phenyl] -d
[Thinyl] -phosphonic acid Preparation of 9A: [2- [2-methylphenyl] -2-oxoethane]-
Phosphonic acid diethyl ester  Dissolve 100 mmol n-butyllithium in 100 ml THF
Of 100 mmol of diethyl methylphosphonate
The stirred solution was added slowly at -78 ° C under argon. Reaction mixture
The mixture was stirred at -78 ° C for 2 hours, dissolved in 50 ml of THF.
50 mmol of o-toluene acid methyl ester
And the reaction mixture at -78 ° C for 20 hours and at 0 ° C for 2 hours
And hydrolyzed with saturated aqueous ammonium chloride.
Normal work-up and flash chromatography on silica gel
45 mmol (90% yield) of the product by chromatography
9A was obtained.

Preparation of 9B: [2- [2-methylphenyl] -ethyl
Nylphosphonic acid diethyl ester 61 mmol (8 ml) of diethylaminosulfur trifluoride (DAS
T) was added slowly at 0 ° C. to a solution of 30 mmol of 9A in 50 ml of anhydrous dichloromethane. The reaction mixture was stirred at 20 ° C. for 30 hours and slowly quenched at 0 ° C. with excess methanol (5 ml). The reaction mixture was evaporated to dryness and purified directly by flash chromatography on silica gel to give 24 mmol of product 9B (80% yield).

[0073]Preparation of 9C: [2- [2-[(2-amino-1,6-dihi
Dro-6-chloro-9H-purin-9-yl) -methyl] -phenyl]-
[Ethynyl] -phosphonic acid diethyl ester  Bromination reaction of 9B and subsequent condensation with 6-chloroguanine
The combination is performed as described for making 1B to 1C.
Was.

[0074]Preparation of 9: [2- [2-[(2-amino-1,6-dihydride
B-6-oxo-9H-purin-9-yl) -methyl] -phenyl] -d
[Thinyl] -phosphonic acid  20 mmol (2.5 ml) of TMSBr is added to 25 ml of anhydrous dichlorometa
A stirred solution of 9C (5 mmol) in argon at 20 ° C under argon
added. The reaction mixture was stirred for 20 hours and evaporated under reduced pressure
Was. The residue was dissolved in 20 ml of acetonitrile,
The body was precipitated by adding 0.5 ml of water. White
The color solid was collected by filtration, 15 ml of 0.2N HCl and 6 ml of
Dissolved in a mixture of THF. Allow this solution at 60 ° C for 8 hours
Heating and the final product 9 is obtained after crystallization by cooling
(1.4 mmol, 28% yield).

[0075]Synthesis of 10: [2- [2-[(2-amino-1,6-dihi
Dro-6-oxo-9H-purin-9-yl) -methyl] -phenyl]-
Ethenyl] -phosphonic acid Preparation of 10A: [2- [2-methylphenyl] -ethenyl] -phos
Phonic acid diethyl ester  38 mmol (1 mole) dissolved in 25 ml of anhydrous tetrahydrofuran
0.95g) bis (diethylphosphonyl) methane slowly
And NaH in anhydrous tetrahydrofuran (42 mmol)
Was added at −15 ° C. under argon. 45 minutes later, 40
38 mmol (4.6 g) dissolved in ml of tetrahydrofuran
Of o-tolualdehyde was added to the reaction mixture at 0 ° C. 20 ℃
After stirring for 18 hours at room temperature, the crude reaction mixture was
Stopped with ammonium and evaporated to dryness. 35 residue
It was suspended in ml of water and extracted three times with 100 ml of ethyl acetate.
The organic layer was washed with brine, dried over sodium sulfate, and filtered.
And evaporated to give 11 g of crude material, which was
Purify by flash chromatography on Kagel 7.
53 g of product 10A were obtained (75% yield).

[0076]Preparation of 10B: [2- [2-[(2-amino-1,6-di
Hydro-6-chloro-9H-purin-9-yl) -methyl] -phenyl
[Ethenyl] -phosphonic acid diethyl ester  Benzoyl peroxide (20mg) in 15ml anhydrous carbon tetrachloride
N-bromosuccinimide (20 mmol) and [2- [2-methyl
Phenyl] -ethenyl] -phosphonic acid diethyl ester (20
mmol). Until all solids rise
At, the mixture was heated to reflux with a lamp. Filter the reaction mixture
And evaporate to give an oil, which is
Dissolved in Lumamide, sodium 6-chloroguanine
Salt (6-chloro guar in 10 ml of anhydrous dimethylformamide)
20 mmol of NaH is added to 20 mmol of NaH at 20 ° C.
(Prepared by addition below). Anti
The reaction mixture was stirred for 20 hours at 20 ° C., evaporated to dryness and the crude residue
The product was purified by flash chromatography on silica gel.
Co-purification to give 12 mmol of product 10B (60% yield)
Was.

[0077]Production of 10: [2- [2-[(2-amino-1,6-dihi
Dro-6-oxo-9H-purin-9-yl) -methyl] -phenyl]-
Ethenyl] -phosphonic acid  Procedure already described for the conversion of 1B to 1 in synthesis 1
Was used to produce 10 from 10B.

[0078]Biological utility  Compounds of the present invention, immunosuppression, anti-lymphoma, anti-leukemia,
As an antiviral and antigenic animal drug, and gout, dry
Acts as a useful drug for the treatment of liquor and autoimmune diseases
Ability is purine nucleotide phosphorylase (PNP)
Can be demonstrated by its ability to inhibit Pudding
Otide phosphorylase (PNP) inhibitory activity was
Can be measured by the bound xanthine oxidase method
It uses inosine as a substrate [HM Calcar,
J. Biol. Chem. 167, 429-443 (1947)]. Apparent dissociation determination
Number (K_I) With 0.1 M HEPES buffer (pH 7.4), 0.05 mM
Four concentrations of inosine ranging from 0.15 mM to
The inhibitor concentration was measured at 1 mM inorganic phosphate. Equation 1
K for a representative member of the compound_iIs shown in Table 1.
Substrate inosine using PNPs from various sources
K_MIs compared to the value of In addition, the compounds of the present invention
Effective against tumors (human MoLT-4 cells)
Have anti-lymphoma and anti-leukemia activity
You. The natural metabolite 2'-deoxyguanosine (about 1 to
10 μM) counteracts lymphoma cells in culture
Seems to be important for the activity to be performed.

K_i(M)PNP source Compound Cow spleen Mouse red blood cells Human red blood cells E. coli  [2- [2-[(2-amino-1,6-dihydro-4x10^-9 2x10^-9 13x10^-9 15x10^-9 6-oxo-9H-purin-9-yl) -methyl] -phenyl] -1,1-difluoroethyl] phosphonic acid [2- [2-[(2-amino-1,6-dihydro-8x10^-Ten 4x10^-Ten 1.8x10^-9 2.5x10^-Ten 6-oxo-9H-purin-9-yl) -methyl] -phenyl] -1-fluoroethenyl] phosphonic acid [2- [2-[(2-amino-1,6-dihydro---3.2x10^-9 5x10^-Ten 6-oxo-9H-purin-9-yl) -methyl] -phenyl] -fluoroethenyl] phosphonic acid [2- [2-[(2-amino-1,6-dihydro-6x10^-Ten 5x10^-Ten 1.3x10^-9 7x10^-Ten 6-oxo-9H-purin-9-yl) -methyl] -phenyl] -1,1,2-trifluoroethyl] phosphonic acid [2- [2-[(2-amino-1,6-dihydro-2.5 x10^-7 3.7x10^-8 2.1x10^-7 − 6-oxo-9H-purin-9-yl) -methyl] -phenoxy] -1,1-difluoropropyl] phosphonic acid [2-[(2-amino-1,6-dihydro-6-7.5 × 10^-9 7.3x10^-9 7.9x10^-8 4.5x10^-9 Oxo-9H-purin-9-yl) -methyl] -phenoxy] -methyl] -phosphonic acid inosine 3x10^-Five 1.5x10^-Four 1.5x10^-Four 8x10^-Five

The term “patient” is used herein in reference to the suppression of the immune system in mice, rats, cats, dogs, cattle,
It shall mean mammals such as sheep, pigs, and primates, including humans. The term "patient" as used herein in the context of treating parasitic infections includes not only mammals, but also other warm-blooded animals such as poultry, including chickens and turkeys.

The term protozoa is intended to include those of the subphylum Sarcomastigophora and Sporozoa of the Protozoa. More specifically, the term protozoa is used herein in humans or human domestic animals.
It is intended to include a genus of parasitic protozoa that is important to humans to cause disease. These genera are most often found in the classification by Baker (1969) as Sarcomastigophora.
It is found to be classified into the subphylum Mastigigophra and the subphylum Sporozoa into the class Telosporea. Representative genera of these parasitic protozoa include Histomonas,
Trypanosoma, Giardia,
Includes Trichomonas, Eimeria, Isopora, Toxoplasma, and Plasmodium.

In fact, a preferred embodiment of the present invention is the use of these compounds as antiprotozoal agents in the treatment of intestinal fluke in poultry on the market. Intestinal fluke (coccidia) infections cause millions of dollars in the poultry industry each year in the United States. Nontoxic and intestinal fluke do not rapidly develop drug resistance due to the rapid development of drug resistance by fluke and due to the relatively high toxicity of some drugs used to treat sporozoosis There is a need for an effective antibacterial agent.

Although the immune system is a major defense mechanism against substances that can cause disease, it cannot distinguish between beneficial and harmful foreign substances and kills both. In many cases, it is beneficial to have a means of controlling the immune system without harm to humans. The compounds of the present invention exhibit such controlling or modulating effects and have potential for use in treating various immune disorders such as rheumatoid arthritis and lupus erythematosus.

The circulating antibodies and the cellular immune response play a role in the rejection of transplanted tissues and organs.
Unless the donor is an identical twin of the recipient, or the individual himself, the recipient's lymphocytes recognize the transplant as non-self and respond immediately to destroy it. An exception to this situation is transplantation into a non-vascularized area (privilege location) such as the cornea of the eye, where lymphocytes do not circulate,
Therefore, it is not sensitized and does not raise an immune response. It is currently difficult to suppress the immune response to prevent transplant rejection without otherwise severely damaging the patient. The patient must also be given large amounts of antibiotics because his own defenses against infection are suppressed. The compounds and compositions of the present invention may be valuable in establishing tolerance for transplantation through controlled modulation of the immune system. Furthermore, these compounds show antiviral activity.

The amount of active ingredient to be administered to a patient can vary widely according to the dosage unit employed, the duration of the treatment, the age and sex of the patient to be treated, the nature and extent of the condition being treated. The total amount of active ingredient to be administered is generally about
The range is 1 mg / kg to 100 mg / kg, preferably 3 mg / kg to 25 mg / kg. A unit dosage form can contain from 25 to 500 mg of the active ingredient and can be taken one or more times per day. The active compounds of the formula I can be administered orally, parenterally or together with a pharmaceutically acceptable carrier in conventional dosage unit form.
Can be administered locally. In a preferred method of administration, 2-deoxyguanosine can be administered with a compound of the present invention. Any non-toxic dosage of 2-deoxyguanosine can be used, typically about 0.5-50 mg / kg per day. For combined use, it is intended to include separate dosage forms as well as those containing 2-deoxyguanosine and a compound of the present invention. The compounds can also be administered in separate dosage units.

The preferred route of administration is oral administration. For oral administration, the compounds can be formulated as solid or liquid preparations, such as capsules, pills, tablets, troches, lozenges, melts,
It can be formulated into a powder, solution, suspension, or emulsion. The solid unit dosage forms can consist of conventional hard or soft shelled gelatins containing surfactants, lubricants and inert fillers, such as lactose, sucrose, calcium phosphate, and corn starch. It may be a capsule. In other embodiments, the compounds of the present invention are combined with common tablet bases such as lactose, sucrose, and corn starch, as well as binders such as gum arabic, corn starch, or gelatin, such as potato starch, alginic acid, corn starch, and guar gum. Disintegrants to assist in breaking and dissolving the tablet after administration, such as talc, stearic acid, or improving the flowability of tablet granules such as magnesium stearate, calcium stearate, or zinc stearate; and tablets for tablet dies and punches Lubricants for preventing the attachment of substances, as well as dyes, coloring and flavoring to enhance the aesthetic properties of the tablet and to make it easier for the patient to consume, can be made into tablets. Suitable excipients for use in oral liquid dosage forms include diluents, such as water and alcohols, with or without pharmaceutically acceptable surfactants, suspending or emulsifying agents. , Such as ethanol, benzyl alcohol, and polyethylene alcohols.

The compounds of the present invention can be injected parenterally, ie, subcutaneously, intravenously, intramuscularly, or intraperitoneally, in a physiologically acceptable diluent containing a pharmaceutical carrier. The carrier may be a pharmaceutically acceptable surfactant such as soap or detergent, for example, a suspending agent such as pectin, carbomer, methylcellulose, hydroxypropylmethylcellulose, or carboxymethylcellulose. Or sterile liquids or mixtures of liquids, with or without emulsifiers and other pharmaceutical adjuvants, such as water, saline, aqueous dextrose and related sugar solutions, alcohols such as ethanol, isopropanol, or Hexadecyl alcohol, glycols such as propylene glycol Are polyethylene glycols, glycerol ketals such as 2,2-dimethyl-1,3-dioxolane-4-methanol, ethers such as poly (ethylene-glycol) 400, oils, fatty acids, fatty acid esters or glycerides, or acetylated Fatty acid glyceride. Examples of oils that can be used in the parenteral formulations of the present invention include those of petroleum, animal, vegetable or synthetic origin,
For example, Nanjing bean oil, soybean oil, sesame oil, cottonseed oil, corn oil, olive oil, petrolatum, and mineral oil.
Suitable fatty acids include oleic, stearic, and isostearic acids. Suitable fatty acid esters are, for example, ethyl oleate and isopropyl myristate. Suitable soaps include fatty acid alkali metal, ammonium, and triethanolamine salts, and suitable detergents include cationic detergents such as dimethyldialkylammonium halides, alkylpyridinium halides, and alkylamines acetates, anions. Detergents, such as alkyl, aryl, and olefin sulfonates, alkyl, olefin, ether, and monoglyceride sulfates,
And sulfosuccinates, nonionic detergents such as aliphatic amine oxides, fatty acid alkanolamides, and polyoxyethylene polypropylene copolymers, and amphoteric detergents such as alkyl-β-aminopropionates, and 2 -Alkylimidazoline quaternary ammonium salts, as well as mixtures. The parenteral compositions of the present invention typically comprise from about 0.5 to about 2 in solution.
Contains 5% by weight of active ingredient. Advantageously, preservatives and buffers are also used. To minimize or eliminate irritation at the injection site, the composition can contain a nonionic surfactant having a hydrophilic-lipophilic balance (HLB) of about 12 to about 17. The amount of surfactant in such formulations ranges from about 5 to about 15% by weight. The surfactant may be a single component having the above HLB, or may be a mixture of two or more components having the desired HLB. Examples of surfactants used in parenteral formulations include polyethylene sorbitan fatty acid esters, such as sorbitan monooleate, and ethylene oxide with a hydrophobic base prepared by propylene oxide / propylene glycol condensation. It is a high molecular weight adduct.

The aerosol or spray compositions containing the compounds according to the invention can be applied to the skin or mucous membranes. Such compositions contain a micronized solid or solution of the compound of formula 1. Also solvents, buffers, surfactants, fragrances,
It may contain antimicrobial agents, antioxidants, and propellants. Such compositions can be applied by means of a propellant under pressure, or by the use of compressible plastic spray bottles, nebulizers or atomizers without the use of gaseous propellants. A preferred aerosol or spray composition is a nasal spray.

The active ingredient may also be released by a sustained release system in which the compound of Formula 1 is slowly released from the inert or biodegradable carrier at a uniform and controlled rate by diffusion, permeation or disintegration of the carrier during the treatment period. Can also be administered. Controlled release drug distribution forms include patches and bandages applied to the skin or sub-lingually or sublingually or intranasally mucosa, intraocular inserts placed in the cul-de-sac of the eye, or gradually administered orally. Tablets or capsules or gastrointestinal reservoirs. Administration by such means of a sustained release distribution system will allow prolonged periods of exposure of bodily tissues to a therapeutically or prophylactically effective dose of a compound of formula 1. A unit dosage of the compound for administration by sustained release system means is approximately the maximum number of days multiplied by the effective daily dose while the carrier remains on or in the body of the host. The sustained release carrier can be in the form of a solid or porous matrix or a reservoir, modified or unmodified cellulose starch,
Gelatin, collagen, rubber, polyolefin, polyamides, polyacrylates, polyalcohols, polyethers, polyesters, polyurethanes, polysulfones, polysiloxanes, polyimides, and polymers of these It can be formed from one or more natural or synthetic polymers, including mixtures and copolymers. The compound of formula 1 can be in pure form in a sustained release carrier or can be dissolved in any suitable liquid or solid vehicle, including the polymer from which the sustained release carrier is formed.

Another aspect of the present invention is to synergize the efficacy of antiviral nucleoside analogs, which would otherwise be subject to the enzymatic action of purine nucleoside phosphorylase.
The use of a purine nucleoside foliphorylase inhibitor of formula 1 in combination therapy.

In particular, the invention encompasses the use of the compounds of formula 1 in combination therapy for the treatment of retroviral infections, especially human infections, and especially for the treatment of human immunodeficiency virus infections. Particularly preferred, 2 ', 3'-dideoxypurine nucleoside is 2', 3'-dideoxyadenosine,
2 ′, 3′-dideoxyguanosine, 2 ′, 3′-dideoxythioinosine and 2 ′, 3′-dideoxyinosine.

The synergistic effect of the antiretroviral effect of, for example, dideoxypurine nucleosides of formula 1 by PNP inhibitors is:
For example, Proc. Nat. Acad. Sci, USA, 83, 1911 (1986).
Can be measured in cell culture media (eg, H9 cells, ATH8 cells) exposed to a retrovirus (eg, HIV) according to methods well known in the art as described in, above. Synergy can also be measured in vivo (eg, in rats) by measuring the increase in dideoxypurine nucleoside plasma levels achieved by prior or concurrent administration of a particular PNP inhibitor according to methods well known in the art. ) Can be measured.

The two active ingredients (2 ', 3'-dideoxypurine nucleoside and PNP inhibitor) can be administered simultaneously by the same or different routes, as the same or different formulations or at different times in time, with the proviso that Provided that an effective PNP inhibitor is present when the 2 ', 3'-dideoxypurine nucleoside is present. The extent to which this temporal separation of the administered active agents can be achieved is:
It depends on the amount of PNP available and the rate at which the PNP inhibitor itself degrades. Preferred dosages for this reason are divided doses two to four times a day, most preferably when both reagents are administered simultaneously.

Purine nucleoside derivatives administered as antiviral agents are catalyzed by purine nucleoside phosphorylase catalysts, which undesirably alter the effects of such agents. In fact, this catalyst can lead to unwanted side effects. Antiviral compounds that are not themselves subject to purine nucleoside phorholase action (eg, if the mechanism is well understood or understood, such as the enzymatic action of enzymatic adenosine deaminase). (If not), it becomes affected by purine nucleoside phosphorylase, and the antiviral effect of such compounds is simultaneously altered. Thus, for the purposes of this aspect of the invention, both antiviral agents are encompassed by the term antiviral agent that receives purine nucleoside phosphorylase.

This aspect of the invention, in another aspect, comprises simultaneously administering a therapeutically effective amount of a purine nucleoside phosphorylase inhibitor, the method comprising the steps of:
Inhibitors, which can be described as a method of treating a viral infection with an antiviral agent that is affected by zeta, are specifically compounds that fall within the general scope of Formula I.

The term antiviral in this aspect of the invention is intended to include the treatment of viruses and diseases caused thereby, which are generally known to be treated with nucleoside analogues, such as AIDS, type B HIV as a causative factor in hepatitis virus and herpes
It contains viruses.

PNPase (purine nucleoside phosphorylase-
Ze) Specific antiviral agents that are currently of interest, with enhanced antiviral effects achieved by combination therapy with inhibitors, are the following compounds. Equation (a)

Embedded imageIn the formula of dideoxynucleosidesR ₁ , R _Two , X name Target virus  OH, H, H dideoxyinosine HIV OH, NH_Two, H dideoxyguanosine HIV, hepatitis B OH, NH_Two, F 3'-F-dideoxyguanosine HIV OH, NH_Two, N_Three 3'-Azidodideoxyguanosine HIV, hepatitis B NH_Two, NH_Two, H dideoxydiaminopurine riboside HIV, HBV NH_Two, H, H dideoxyadenosine HIV NH_Two, NH_Two, N_Three 3'-Azidodideoxydiaminopurine riboside HIV, HBV The last three compounds are the first of adenosine deaminase
Substrate. Equation (b)

Embedded imageIn the formula of dideoxydehydronucleosidesR ₁ , R _Two , Name Target virus  OH, H, dideoxydehydroinosine HIV OH, NH_Two, Dideoxydehydroguanosine HIV NH_Two, N dideoxydehydroadenine HIV (c) formula

Embedded image R ₁ and R _{2 in the} formula of the dioxolamprin derivative are changed,
R ₁ is H or NH ₂ , R ₂ is H or NH ₂ , the compound is a target for HIV and HBV (ie, hepatitis B), and (d)

Embedded image Wherein R ₁ is OH or NH ₂ , and R ₂ is H or NH ₂ . The target virus is HIV.

It should be noted that currently the two best antivirals against hepatitis B (HBV) are dideoxyguanosine (ddGuo) and 2,6-diiminodideoxypurine riboside, a prodrug of dideoxyguanosine. It is.

Particularly useful compounds of the invention are those of the formula

Embedded image Wherein R ₃ is H, Ar is 2,3-thiophene, 2,5-furan or 3,4-furan, and Z ″ is —CH ₂ CF ₂ −
P (OH) ₂ , Ar is 1,2-phenyl, R ₃ is H or NH ₂ , Z ″ is CH ₂ CF ₂ PO (OH) ₂ , CH ₂ CHFPO (OH) ₂ , C
HFCF ₂ PO (OH) ₂ , CHOHCF ₂ PO (OH) ₂ , CH = CFPO (OH) ₂ , CH = CHP
O (OH) ₂ or COCF ₂ PO (OH) ₂ . As with most classes of chemotherapeutic agents, certain subgeneric and certain specific embodiments exhibit more beneficial profiles than others. Preferred compounds in the PNP inhibitors of Formula 1 are those wherein R ₁ is OH, R ₂ is NH ₂ , R ₃ is H or NH ₂ , Ar is 1,2-phenyl or 2,3-furan or thiophene, R ₅ and R ₆ is a compound of H. Preferred Z moieties are (a) X and Y are F, R ₄ is H, R ′ ₄ is H or F,
(b) n is 0, X and Y are both H, (c) X is F, Y is F or H, (d) X is F, and Y is F Or (e) X and Y are F,
R ₇ and R ₈ are H. Preferred specific compounds are the end products of Examples 1 to 10 as well as the amino analogue in the 3-position.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁷ Identification code FI A61K 31/675 A61K 31/675 (72) Inventor Charles Danzine France Strasbourg 67,000 Le Geirer 18 (58) Fields surveyed ( Int.Cl. ⁷ , DB name) C07F 9/6561 CA (STN) REGISTRY (STN)

Claims (17)

(57) [Claims] (1) Formula (1) [Wherein R is Wherein Ar is a group wherein the CH ₂ moiety next to Ar is attached to one ring carbon atom of the R ₉ substituted phenyl, R ₉ substituted thiophene or R ₉ substituted furan moiety, and Z moiety adjacent to Ar Is a bridging moiety attached to the second ring carbon atom of the R ₉ -substituted phenyl, R ₉ -substituted thiophene or R ₉ -substituted furan moiety; and Z is the following (a), (b), (c), (d) or (e), and (a)
Is And (b) is And (c) is (d) is -C≡C-, and (e) is With the proviso that when Z is part (b), Ar is other than a furan or thiophene part, and n is 1
An integer of 0 or 0; R ₁ is —OH or —SH; R ₂ is H or —NH ₂ ; R ₃ is H, —NH ₂ , —OH or —NH—NH ₂ R ₄ is H, R ′ ₄ is H, OH, or F, or R ₄ and R ′ ₄ together form a keto moiety together with the carbon atom to which they are attached; ₅ is C _1-6 alkyl or R ′ ₅ , R ₆ is C _1-6 alkyl or R ′ ₆ , R ′ ₅ and R ′ ₆ are H, and each R ₇ and R ₈ is H , F, or C _1-4 alkyl; and R ₉ is H, Cl, Br, C _1-6 alkyl, C _1-6 alkoxy, OH, NH ₂ or CH ₃ , provided that Ar is furan or thiophene. Where R ₉ is other than OH or NH ₂ , and X and Y are H, F, or Cl, provided that when n is 0, both X and Y are H On condition that That. A compound thereof, a tautomeric compound thereof, and a pharmaceutically acceptable salt thereof. 2. The compound according to claim 1, wherein R ₅ and R ₆ are H. 3. The compound according to claim 1, wherein R ₁ is OH. 4. The compound according to claim 1, wherein R ₂ is NH ₂ . 5. The compound according to claim 1, wherein R ₃ is H. 6. The compound according to claim 1, wherein R ₃ is NH ₂ . 7. The compound according to claim 1, wherein Ar is phenyl or thiophene. 8. Z is CHFCF ₂ , CH = CF or CH
2. The compound according to claim 1, wherein = CH. 9. R ₁ is OH, R ₂ is NH ₂ ,
R ₃ is H, Ar is phenyl, and Z is —CH ₂ C
_{F 2, -CH 2 CHF, -CHFCF} 2, CH (OH) C
_{F 2, -C (O) CF} 2, -CH = CF, and -CH = C
The compound according to claim 2, which is selected from the group consisting of H. 10. R ₁ is OH, R ₂ is NH ₂ , R ₃ is NH ₂ , Ar is phenyl, and Z is-
_{CH 2 CF 2, -CH 2 CHF} , -CHFCF 2, CH (O
_{H) CF 2, -C (O} ) CF 2, -CH = CF, and -C
3. The compound according to claim 2, wherein the compound is selected from the group consisting of H = CH. [11] [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl) -methyl] -phenyl] -1,1-difluoroethyl] phosphone Acid [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purine-9
-Yl) -methyl] -phenyl] -1-fluoroethenyl] phosphonic acid [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purine-9)
-Yl) -methyl] -phenyl] -1,1,2-trifluoroethyl]
Phosphonic acid [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purine-9
-Yl) -methyl] -phenyl] -2-hydroxy-1,1-difluoroethyl] phosphonic acid [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purine-9)
-Yl) methyl] -phenyl] -2,2-dihydroxy-1,1-difluoroethyl] phosphonic acid [3- [2-[(2-amino-1,6-dihydro-6-chloro-9H-purine- 9
-Yl) methyl] -phenoxy] -1,1-difluoropropyl]
Phosphonic acid [2-[(2-amino-1,6-dihydro-6-oxo-9H-purin-9-yl) methyl] -phenoxy] methylphosphonic acid [2- [2-[(2-amino-1, 6-dihydro-6-oxo-9H-purine-9
-Yl) methyl] -phenyl] -1,1-difluoro-2-propenyl] phosphonic acid [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purine-9)
-Yl) methyl] -phenyl] ethynyl] phosphonic acid [2- [2-[(2-amino-1,6-dihydro-6-oxo-9H-purine-9)
The compound according to claim 1, which is a compound selected from the group consisting of -yl) methyl] -phenyl] ethenyl] phosphonic acid. 12. A therapeutic agent for protozoan infection, comprising a therapeutically effective amount of the compound of claim 1. 13. An agent for treating a viral infection, comprising a therapeutically effective amount of the compound according to claim 1. 14. A therapeutic agent for gout, comprising a therapeutically effective amount of the compound of claim 1. 15. An agent for treating a viral infection, which comprises a therapeutically effective amount of the compound according to claim 1 and is administered in combination with 2 ′, 3′-dideoxypurine nucleoside. 16. An agent for treating retroviral infection, which comprises a therapeutically effective amount of the compound of claim 1 and is administered in combination with 2 ', 3'-dideoxypurine nucleoside. 17. A compound of the formula Wherein R ′ is —CH ₂ —Ar—Z—P (O) (OH) ₂ , wherein Ar is a group in which the CH ₂ portion next to Ar is R ₉ -substituted phenyl, R ₉ -substituted thiophene or R ₉ The Z moiety adjacent to Ar is attached to one ring carbon atom of the _9- substituted furan moiety and the Z moiety adjacent to Ar is attached to the second ring carbon atom of the R ₉ -substituted phenyl, R ₉ -substituted thiophene, or R ₉ -substituted furan moiety Z is the following part of (a), (b), (c), (d), or (e), and (a)
Is And (b) is And (c) is (d) is -C≡C-, and (e) is Provided that when Z is a moiety (b), Ar is other than a furan or thiophene moiety, n is an integer of 1 to 5 or 0, and R ₁ is -OH or -SH and a, R ₂ is H or -NH _2, R ₃ is H, -NH _2, -OH or -NH-NH _2, R ₄ is H, R _'4 is, H, OH, Or R, or R ₄ and R ′ ₄ together form a keto moiety with the carbon atom to which they are attached, and each R ₇ and R ₈ is H, F, or C _1-4 alkyl. R ₉ is H, Cl, Br, C _1-6 alkyl, C _1-6 alkoxy, OH, NH ₂ or CH ₃ , provided that when Ar is furan or thiophene, R ₉ is OH Or X and Y are H, F, or Cl provided that they are other than NH ₂ , provided that when n is 0, X and Y are both H. Wherein the compound is selected from the group consisting of: a) tautomeric compounds thereof, and pharmaceutically acceptable salts thereof. [Wherein R ″ is A) contacting the compound with trimethylsilyl bromide under standard reaction conditions, and optionally converting the product to a pharmaceutically acceptable salt.